3D tessellation imaging

Abstract

The invention provides a new system and method for imaging a specimen. The system projects a three-dimensional crystalline pattern of light, a tessellation, and records the specimen's emitted light at locations where a portion of the specimen coincides with the pattern.

We claim: 
     
         1 . A system for imaging a specimen comprising:
 a plurality of radiation beams   directing means to direct said plurality of radiation beams in a beam geometry designed to produce a crystalline pattern of radiation intensity in a volume of interference   positioning means to position said specimen such that at least a portion of the specimen coincides with said volume of interference   imaging means to image radiation emitted from said specimen in response to said pattern   
     
     
         2 . The system of  claim 1  wherein said pattern is substantially stationary. 
     
     
         3 . The system of  claim 1  further comprising a beam modulator constructed to modulate at least one of said plurality of radiation beams to control said pattern. 
     
     
         4 . The system of  claim 1  wherein said emitted radiation is fluorescence. 
     
     
         5 . The system of  claim 1  wherein said imaging means comprises a first camera focused on a first plane within said pattern. 
     
     
         6 . The system of  claim 5  wherein said imaging means further comprises a second camera focused on a second plane within said pattern. 
     
     
         7 . The system of  claim 1  wherein said imaging means further comprises at least one objective lens. 
     
     
         8 . The system of  claim 7  wherein at least one of said plurality of beams emerges from said at least one objective lens. 
     
     
         9 . The system of  claim 1  wherein said pattern comprises a plurality of intensity peaks. 
     
     
         10 . The system of  claim 9  wherein said imaging means comprises a collection point spread function substantially aligned with one of said peaks. 
     
     
         11 . A method for imaging a specimen comprising:
 providing a plurality of radiation beams   directing said plurality of beams in a beam geometry designed to produce a crystalline pattern of radiation intensity in a volume of interference   positioning said specimen such that at least a portion of the specimen coincides with said volume of interference   imaging radiation emitted from said specimen in response to said pattern   
     
     
         12 . The method of  claim 11  wherein said positioning further comprises scanning said specimen relative to said volume. 
     
     
         13 . The method of  claim 12  wherein said imaging further comprises recording a raw data set of emitted radiation intensity at a plurality of positions during said scanning. 
     
     
         14 . The method of  claim 13  further comprising estimating specimen brightness from said raw data set by numerical deconvolution of a type chosen from the list including linear, nonlinear, iterative, or a combination of these. 
     
     
         15 . The method of  claim 11  wherein said directing further comprises controlling beam parameters chosen from the list including angle, phase, intensity, polarization, wavelength. 
     
     
         16 . The method of  claim 15  wherein said controlling controls said pattern to be substantially stationary. 
     
     
         17 . The method of  claim 15  wherein said controlling controls said pattern to change over time.